Super-resolution microscopy of nuclear lamin and spindle envelope/matrix function

核纤层和纺锤体包膜/基质功能的超分辨率显微镜

• 批准号: 8666780
• 负责人: ROBERT D GOLDMAN
• 金额: $ 28.94万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8666780
• 关键词: 4D Imaging; Address; Animals; Area; Attention; Behavior; Bulla; Cell Cycle; Cell Nucleus; Cell Proliferation; Cell division; Cells; Cellular Structures; Cellular biology; Chromatin; Data; Dynein ATPase; Ensure; Human; Interphase; Knockout Mice; Laboratories; Lamin Type B; Lamins; Lighting; Malignant Neoplasms; Mechanics; Membrane; Membrane Proteins; Microscopy; Microtubules; Mitosis; Mitotic; Mitotic spindle; Molecular; Morphogenesis; Mutation; Nuclear Envelope; Nuclear Lamin; Nuclear Lamina; Nuclear Pore; Nuclear Pore Complex; Nuclear Proteins; Optics; Patients; Prophase; Protein Isoforms; Resolution; Structure; Techniques; daughter cell; embryonic stem cell; human disease; insight; light microscopy; link protein; new therapeutic target; physical model; public health relevance; reconstruction; three dimensional structure

DESCRIPTION (provided by applicant): This proposal aims to study the structural organization of the different types of lamin networks/meshworks during interphase and mitosis using super resolution microscopy techniques. In Aim 1, we plan to determine whether each lamin (LA, LC, LB1, LB2) forms a separate lamina meshwork or whether the establishment of a lamina meshwork requires the interactions of different lamin isoforms in normal interphase nuclei. We will also study the interactions of each of the lamins with chromatin and nuclear pore complexes (NPCs). As animal cells enter mitosis, parts of the interphase membrane network, including the nuclear lamina, and nuclear proteins re-organize into an inter-connected network that both surrounds and permeates the spindle microtubules. This network has been referred to as either the spindle envelope (or sheath) or spindle matrix. Here we will refer to the network as the Spindle envelope/matrix (Spenix). The major challenge in defining the function of the Spenix is to visualize the behavior of Spenix proteins and link their behaviors to their functions in mitosis. In Aim 2, we will apply super resolution microscopy to study the membrane and protein organization of the Spenix, which should open the door to dissect not only the organization of the Spenix but also the functional relationship of the Spenix and the mitotic spindle. The results from the proposed studies will provide important new information regarding the interactions and functions of the nuclear lamins and their interactions with their major interacting partners in interphase and mitosis.

描述（由申请人提供）：本提案旨在使用超分辨率显微镜技术研究间期和有丝分裂期间不同类型核纤层网络/网状结构的结构组织。在目标 1 中，我们计划确定每个核纤层蛋白（LA、LC、LB1、LB2）是否形成单独的核纤层网络，或者核纤层网络的建立是否需要正常间期核中不同核纤层蛋白亚型的相互作用。我们还将研究每个核纤层蛋白与染色质和核孔复合物 (NPC) 的相互作用。当动物细胞进入有丝分裂时，部分间期膜网络（包括核纤层）和核蛋白重新组织成一个相互连接的网络，围绕并渗透纺锤体微管。该网络被称为纺锤体包膜（或鞘）或纺锤体基质。在这里，我们将网络称为主轴包络/矩阵（Spenix）。定义 Spenix 功能的主要挑战是可视化 Spenix 蛋白的行为并将其行为与其有丝分裂中的功能联系起来。在目标 2 中，我们将应用超分辨率显微镜来研究 Spenix 的膜和蛋白质组织，这将为解剖 Spenix 的组织以及 Spenix 和有丝分裂纺锤体的功能关系打开大门。拟议研究的结果将提供有关核纤层蛋白的相互作用和功能以及它们与间期和有丝分裂中主要相互作用伙伴的相互作用的重要新信息。